骨髓间充质干细胞不同时相干预对SD大鼠急性肺损伤的修复作用

目的 探究骨髓间充质干细胞(BM-MSCs)不同时间干预对以静脉注射脂多糖(LPS)构建大鼠急性肺损伤(ALI)模型的修复作用.方法 体外分离、培养并鉴定大鼠BM-MSCs.将120只雄性SD大鼠纳入研究,分成对照组、LPS组和LPS+ BM-MSCs组,上述3组因BM-MSCs处理时间差异分成2、8、24、48、96 h共5个亚组(n=8).LPS组以5 mg/kg尾静脉注射LPS,LPS+ BM-MSCs组在注射LPS后分别在上述不同时间点尾静脉注射1 mL的BM-MSCs(1×106·mL-1);对照组则在上述时相点注射等剂量的生理盐水.检测各组肺泡灌洗液肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1α等炎症因子浓度、肺湿干比及肺损伤病理评分.结果 成功分离、培养、纯化BM-MSCs,并诱导向成骨、成软骨、成脂细胞分化;流式细胞术鉴定所培养细胞系CD90、CD105、CD45、CD34阳性率分别为98.89％、97.37％、3.17％及1.41％.在LPS诱导·的ALI中,TNF-α、IL-1α、IL-10在8h亚组中测得最高值[分别为(364.911±32.182) pg/mL、(286.316±18.567) pg/mL、(162.037±22.377) pg/mL],随着时间延长逐渐下降;肺湿干比在2h亚组中测得最高值(6.361 ±0.265);肺水肿及组织损伤程度在2h亚组中最严重,病理评分也测得最高值(0.462±0.048),随着时间延长逐渐下降.在LPS+ BM-MSCs组中,2、8、24 h的炎症因子、湿干比及病理评分均有显著改变(P＜0.05),随着时间延长,差异无统计学意义(P＞0.05).结论 早期应用BM-MSCs可减少TNF-α、IL-1α等炎症因子,从而减轻肺组织损伤及肺泡水肿.

The effects of bone marrow mesenchymal stem cells intervention at different times on acute lung injury repairmen

Objective To investigate the intervention effects of bone marrow-derived mesenchymal stem cells (BM-MSCs) on the repairmen of acute lung injury (ALI) at different times.Methods The rat BM-MSCs were isolated,cultured and identified in vitro.120 male SD rats were divided into control group,LPS group and LPS + MSCs group (The above three groups were divided into 5 subgroups (2 h,8 h,24 h,48 h,96 h,n =8) according to different time points of MSCs administration);in which placebo saline,lipopolysaccharide (LPS),and LPS plus MSCs treatment were given,respectively.The inflammation factors,including TNF-α,IL-1 α and et al.,in bronchoalveolar lavage fluid,the lung wet/dry ratio,and pathological score of lung injury were compared.Results With the cells isolated,cultured and further purified,the BM-MSCs can be induced to differentiate to adipocyte-oriented,chrondrocyte-oriented and osteocyte-oriented cells.The positive rates of CD90,CD105,CD45 and CD34 were respectively 98.89％,97.37％,3.17％ and 1.41％ by flow cytometry.The maximum concentration of TNF-α,IL-1α,IL-10 were observed in 8 h subgroup of LPS group [(364.911 ± 32.182) pg/mL,(286.316 ± 18.567) pg/mL,(162.037 ± 22.377) pg/mL],and gradually decreased over time,while the maximum lung wet/dry ratio and pathological grading were observed in 2 h subgroup of LPS group(6.361 ±0.265,0.462 ± 0.048),then gradually decreased over time.As early intervention (2 h,8 h,24 h) of BM-MSCs,the inflammatory factor,dry wet ratio and pathological score were significantly different(P ＜0.05),while there were no significant difference over time (P ＞ 0.05).Conclusion Early application of BM-MSCs can reduce TNF-α and IL-1 α,so as to reduce the lung injury and alveolar edema.